Figure 6
The production of mature LHCGR isoforms in chorinic villi from Down's syndrome pregnancies are significantly reduced compared to that of controls. The CVS samples were Tri-zol extracted to recover mRNA as well as proteins. Approximately 10 μg of total protein was loaded in each lane. The proteins extracted from DS (T21) CVS (a) and control CVS (b) pregnancies were resolved in 8% polyacrylamide-SDS gels, Western blotted and immunoreacted with anti-human LHCGR (LHR-29) monoclonal antibody. Blots were stripped prior to immunostaining with anti-β-Actin monoclonal antibody. The data shown in a) and b) were from the same experiment except that the control and DS proteins were separated in two gels at the same time. In order to compare the band intensity in different experiments, two known CVS samples in duplicate were incorporated in each experiment. The density of the 80K LHCGR and 42K β-actin bands served as references for quantitative analysis of the experimental samples. The relative migration of the isoforms is indicated by an arrow. The Mr 80K protein band (LHCGR p80), indicated by * in a) and b), well separated from the neighboring variants were scanned and c) the relative densities of the LHCGR p80 with respect to β-Actin in normal and trisomic pregnancies, n = total number of experiments carried out on protein samples in each condition. **P < 0.01.